PRESS-NEWS.org - Press Release Distribution
PRESS RELEASES DISTRIBUTION

MGH researchers develop faster method of engineering zinc-finger nucleases

Less labor-intensive approach speeds creation of powerful DNA-targeting tool

2010-12-12
(Press-News.org) A team led by Massachusetts General Hospital (MGH) researchers has developed a faster way to engineer synthetic enzymes that target specific DNA sequences for inactivation, repair or alteration. The report from the MGH Molecular Pathology Unit, being published online in Nature Methods, describes a highly effective but less labor-intensive way to generate powerful tools called zinc-finger nucleases (ZFNs).

"With our approach, called context-dependent assembly, any scientist can use either standard molecular biology techniques or commercial DNA synthesis to design ZFNs for their target gene of interest," says J. Keith Joung, MD, PhD, associate chief for Research in MGH Pathology, the study's senior author. "ZFNs are broadly applicable, powerful tools for manipulating the genomes of cells from various organisms – including humans – and may provide a way to efficiently correct gene mutations responsible for human disease, avoiding problems resulting from the imprecise nature of current gene therapy approaches using viral vectors."

Most human transcription factors that control whether a genetic signal is translated into a protein bind to specific DNA sequences using peptides called zinc fingers. Zinc-finger nucleases are synthetic "designer" proteins combining a zinc-finger domain, engineered to bind a particular DNA sequence, with an enzyme that breaks both DNA strands at the targeted site. While ZFNs have great potential, creating the customized proteins has been challenging.

In the simplest approach, called modular assembly, individual peptides are linked together like beads on a string to create a multi-finger protein theoretically able to recognize long DNA segments. Joung and others have shown that, in practice, modular assembly has a very low success rate for creating multi-finger proteins. This high failure rate is most likely due to "context-dependent" effects that individual zinc fingers can have on the DNA-binding activities of their neighboring fingers. Assembling peptides that don't work well together would be like trying to put together jigsaw puzzle pieces that don't fit.

In 2008, Joung and colleagues at the University of Minnesota and other institutions, members of the Zinc Finger Consortium, reported developing a method called OPEN (Oligomerized Pool ENgineering), which takes these context-dependent effects into account. But although OPEN works well, it can be labor intensive and extremely time consuming – requiring up to a year for a lab to establish the technology and two months of work to generate desired ZFNs. To address these limitations, the MGH research team has assembled an extensive archive of zinc fingers known to work well when positioned together – in essence puzzle pieces that already have been put together. Using this context-dependent method, the investigators were able to assemble dozens of ZFNs in as little as four days.

"With this archive in hand, any researcher can easily generate their own ZFNs in less than a week, and no special expertise is needed," Joung explains. "In addition to being much faster, context-dependent assembly can generate large numbers of ZFNs simultaneously, which is hard to do with OPEN because it is more labor intensive." As was the case with OPEN, the Joung lab and the Zinc Finger Consortium (http://www.zincfingers.org) will make the software and reagents required to practice context-dependent assembly available to all academic laboratories.

"One of the holy grails of genetics is the ability to make targeted changes to individual genes," says Laurie Tompkins, PhD, who over sees genetics grants at the National Institute of General Medical Sciences, one of the National Institutes of Health and a major supporter of this study. "Dr. Joung and his colleagues have developed an extraordinarily simple, efficient strategy for using zinc finger technology to swap out altered versions of genes for normal ones – or vice versa – providing basic scientists and clinicians alike with a broadly applicable research tool."

Adds Joung, an associate professor of Pathology at Harvard Medical School, "At this point, I believe that context-dependent assembly will have the biggest impact on researchers using ZFNs to genetically manipulate model organisms, possibly even models developed from pluripotent stem cells. Other big impacts should be enabling researchers to create knockout mutations in a large series of genes involved in a common pathway or related to a specific disease and to use ZFNs to create comprehensive collections of mutants for every gene in an organism." Joung is also a member of the MGH Center for Computational and Integrative Biology and Center for Cancer Research

The challenges posed to scientists interested in using ZFNs in their investigations were described in an article in the Fall 2010 issue of the MGH-sponsored magazine Proto, which can be accessed at http://protomag.com/assets/zinc-fingers-entry-fee.

INFORMATION:

Jeffry Sander, PhD, of the MGH Molecular Pathology Unit is lead author of the Nature Methods report. Additional co-authors are Elizabeth Dahlborg, Mathew Goodwin, Jessica Blackburn, Stacey Thibodeau-Beganny, Morgan Maeder, Cyd Khayter and David Langenau, MGH Molecular Pathology; Lindsay Cade, Randall Peterson and Jing-Ruey Yeh, MGH Cardiovascular Research Center; Feng Zhang, Shaun Curtin, Yiping Qi, Christopher Pierick, Robert Stupar and Daniel Voytas, University of Minnesota; Daniel Cifuentes, Ellen Hoffman and Antonio Giraldez, Yale University; and Deepak Reyon and Drena Dobbs, Iowa State University. Support for the study includes grants from the National Institutes of Health and the National Science Foundation.

Massachusetts General Hospital, established in 1811, is the original and largest teaching hospital of Harvard Medical School. The MGH conducts the largest hospital-based research program in the United States, with an annual research budget of more than $600 million and major research centers in AIDS, cardiovascular research, cancer, computational and integrative biology, cutaneous biology, human genetics, medical imaging, neurodegenerative disorders, regenerative medicine, systems biology, transplantation biology and photomedicine.

END



ELSE PRESS RELEASES FROM THIS DATE:

Protein offers new clue to cause and treatment for kidney disease

2010-12-12
BIRMINGHAM, Ala. – University of Alabama at Birmingham researchers have pinpointed a protein that compromises the kidney's filtering ability, causing nephrotic syndrome, and demonstrated that a naturally occurring precursor of an acid in the body offers potential for treating some forms of the condition. The research was published online Dec. 12 in Nature Medicine. "This is a major breakthrough in understanding the development and treatment of kidney disease associated with proteinuria, the leakage of protein in the urine," said the study's lead author Sumant Singh ...

UCR scientists identify pomegranate juice components that could stop cancer from spreading

UCR scientists identify pomegranate juice components that could stop cancer from spreading
2010-12-12
RIVERSIDE, Calif. – Researchers at the University of California, Riverside have identified components in pomegranate juice that both inhibit the movement of cancer cells and weaken their attraction to a chemical signal that promotes the metastasis of prostate cancer to the bone. The research could lead to new therapies for preventing cancer metastasis. Performed in the lab of Manuela Martins-Green, a professor of cell biology, the research was presented today (Dec. 12, 2010) at the 50th annual meeting of the American Society for Cell Biology taking place in Philadelphia. Prostate ...

John Wayne Collectables Expands to Meet Growing Demand

2010-12-12
The first two added products are seasonal and will not be available long, a 2011 calendar and a Christmas ornament. The third added product is a new figurine display called the Sunset Rider. Behind the hand-sculpted resin figure is a display with a blazing western sky, a portrait of "The Duke", and an image of John Wayne riding his horse. The border of the display is crafted to look like stone. The fourth new item is a set of four glasses, six inches high, and each glass holding ten ounces. This set has two designs, each with a portrait of John Wayne on the front, ...

Strategic Analysts Publishes an In Depth Analysis of North American Newsprint Markets

2010-12-12
Demand for newsprint in North American has declined by 50% over the past decade, and despite recent price increases the future remains bleak for producers. Strategic Analysts has released their latest white paper discussing the decline and analyzing the cost competitiveness and financial viability of AbitibiBowater and the other major producers. Springboro, OH - December 3, 2010 - Back in 2000, newsprint demand in North America was at an all time high. Daily circulation was at its record levels and the newspapers themselves were reporting record advertising revenue. ...

Pizza.com Signs Shay Traynor to the Sales Team

2010-12-12
California-based Pizza.com has announced the addition of Shay Traynor to the company's sales staff. Ms. Traynor comes to Pizza.com from Restaurant.com, an online provider of discounted dining gift certificates that helps independent restaurants promote aspects of their business. There, she received recognition for being a top ten representative and with 15+ years of experience in the restaurant industry, was able to connect with restaurant owners and expand Restaurant.com's territory. She has previously served as a restaurant manager at The MilkyWay Restaurant and Tapas ...

Emerson Bearing Promotes Bob Piracini to Marketing Specialist to Lead New Machine Tools Division

2010-12-12
As Marketing Specialist, Piracini will lead the new Machine Tools division and assist clients in determining which bearing technologies would be most suitable by taking into consideration the specifications, recommendations, maintenance strategies, fatigue life and wear resistance of the bearing in relation to the application. Piracini's clients are typically spindle repair and machinery rebuilds. They range from regional and national "mom and pop" operations to large companies. Piracini will continue to travel throughout New England several times a year visiting companies ...

Niche Website LightBoxes4Sale.com Ups Light Box Offering

2010-12-12
Access Display Group, Inc. (ADG) has launched a new B2B e-commerce website offering light boxes in a wide range of styles, finishes and standard and custom sizes. LightBoxes4Sale.com is a niche website that focuses on a very specific sector of the display fixture market: light boxes. This narrow scope means more specialized, targeted customer care and a wider selection. Explains ADG sales and marketing director Craig Abrams, "If you're a designer or corporate buyer and you are looking for a light box, LightBoxes4Sale.com will have the perfect style and size for you." Light ...

Lisa Calkins, CEO of Amadeus Consulting, Nominated as Most Influential Women in Technology

2010-12-12
Lisa Calkins, CEO of custom application development company Amadeus Consulting is nominated as one of the most influential women in tech for 2011. The awards are put on by Fast Company, a magazine that is at the forefront of exciting emerging technology and innovation in growing companies. The annual award is given to female power players in the technology industry who bring a unique brand thought leadership and proven success to technology companies in all sectors including: custom software development, consumer product, mobile application development, etc. The award ...

Fishbowl Solutions Releases Enterprise Batch Loader for Oracle Universal Content Management 11g

2010-12-12
News Facts • Fishbowl Solutions, a consulting services and software development company specializing in solutions for Oracle's Universal Content Management and WebCenter products, has released Enterprise Batch Loader, a software application that provides an efficient and powerful method to check in file and data formats generated by third-party systems to Oracle Content Server. • According to AIIM (Association for Information and Image Management), 80% of an organization's information is unstructured and 90% of this remains unmanaged and continues to grow at roughly ...

U.S. Computer Desk Retails Offer Huge Discounts on Mayline Furniture

2010-12-12
For a limited time, ComputerDesksNMore.com is offering up to 60% off on select Mayline Group products. This end of year close out sale wonâEUR t last forever, and you can select from a number of different Mayline products, including bookcases, computer desks, conference tables, filing cabinets and other office and home furniture. Established in 1939, Mayline has become a market leader in office ergonomics, offering the marketâEUR s largest and diverse selection of office materials. At ComputerDesksNMore, youâEUR ll find the best selection of Mayline furniture at competitive ...

LAST 30 PRESS RELEASES:

Study unexpectedly finds living in rural, rather than urban environments in first five years of life could be a risk factor for developing type 1 diabetes

Editorial urges deeper focus on heart-lung interactions in pulmonary vascular disease

Five University of Tennessee faculty receive Fulbright Awards

5 advances to protect water sources, availability

OU Scholar awarded Fulbright for Soviet cinema research

Brain might become target of new type 1 diabetes treatments

‘Shore Wars:’ New research aims to resolve coastal conflict between oysters and mangroves, aiding restoration efforts

Why do symptoms linger in some people after an infection? A conversation on post-acute infection syndromes

Study reveals hidden drivers of asthma flare-ups in children

Physicists decode mysterious membrane behavior

New insights about brain receptor may pave way for next-gen mental health drugs

Melanoma ‘sat-nav’ discovery could help curb metastasis

When immune commanders misfire: new insights into rheumatoid arthritis inflammation

SFU researchers develop a new tool that brings blender-like lighting control to any photograph

Pups in tow, Yellowstone-area wolves trek long distances to stay near prey

AI breakthrough unlocks 'new' materials to replace lithium-ion batteries

Making molecules make sense: A regional explanation method reveals structure–property relationships

Partisan hostility, not just policy, drives U.S. protests

The Journal of Nuclear Medicine Ahead-of-Print Tip Sheet: August 1, 2025

Young human blood serum factors show potential to rejuvenate skin through bone marrow

Large language models reshape the future of task planning

Narrower coverage of MS drugs tied to higher relapse risk

Researchers harness AI-powered protein design to enhance T-cell based immunotherapies

Smartphone engagement during school hours among US youths

Online reviews of health care facilities

MS may begin far earlier than previously thought

New AI tool learns to read medical images with far less data

Announcing XPRIZE Healthspan as Tier 5 Sponsor of ARDD 2025

Announcing Immortal Dragons as Tier 4 Sponsor of ARDD 2025

Reporting guideline for chatbot health advice studies

[Press-News.org] MGH researchers develop faster method of engineering zinc-finger nucleases
Less labor-intensive approach speeds creation of powerful DNA-targeting tool